Two aspects of the technique of affinity chromatography (AC) will be studied. We propose to develop methods and materials to permit high speed analytical affinity chromatography (HSAAC) to become a reality. Second, we propose to study the nature of the thermodynamic and kinetic processes which are involved in conventional affinity chromatography. In order to carry out HSAAC we will study, prepare and characterize the chromatographic properties of various glasses, silicas, chemically modified silicas, and other hydrophilic solids. Thermal pre-treatment of the solids may be able to minimize the number of nonspecific protein adsorption sites. Glasses will be modified by a variety of silanization agents, judiciously chosen to minimize both residual Si-OH groups on the solid and additional Si-OH groups introduced by the use of trialkoxy (or trichloro) silanes. End-capping with non-ionic, non-adsorptive silanes will be investigated. Our objective will be to generate a micro particle (5-10 micron) solid support which is optimized for three critical chromatographic factors: efficiency, selectivity, and capacity. These supports will be used with a general affinity ligand (AMP). We will characterize these affinity sorbents via the first four peak moments (area, centroid, variance, asymmetry) as a function of sample size and flow rate. The second study proposes to examine the thermodynamic and kinetic properties of conventional affinity chromatographic systems. We will carry out studies of the effect of solid phase, pore size, spacer arm length, pH, ionic strength, temperature, and bio-selective eluting agents on the retention volume and linear binding capacity of affinity sorbents. The focus of these studies will be the elucidation of the vital spacer arm length effect. We will carry out our studies by a variety of complementary chromatographic, calorimetric and equilibrium dialysis methods. We will compare binding constants measuresd for both solution phase and solid phase reactions so as to define the role played by the affinity matrix.